WO2008147296A1 - Procédé de traitement enzymatique d'une boue dans la purification de l'eau - Google Patents

Procédé de traitement enzymatique d'une boue dans la purification de l'eau Download PDF

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Publication number
WO2008147296A1
WO2008147296A1 PCT/SE2008/000376 SE2008000376W WO2008147296A1 WO 2008147296 A1 WO2008147296 A1 WO 2008147296A1 SE 2008000376 W SE2008000376 W SE 2008000376W WO 2008147296 A1 WO2008147296 A1 WO 2008147296A1
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WIPO (PCT)
Prior art keywords
sludge
enzymes
enzyme
bacteria
chosen
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Application number
PCT/SE2008/000376
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English (en)
Inventor
Estera Szwajcer Dey
Olof NORRLÖW
Original Assignee
Kemira Oyj
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Kemira Oyj filed Critical Kemira Oyj
Priority to AU2008257660A priority Critical patent/AU2008257660A1/en
Priority to CN2008800178774A priority patent/CN101679089B/zh
Priority to EP08767049A priority patent/EP2158166B1/fr
Priority to BRPI0810931-1A2A priority patent/BRPI0810931A2/pt
Priority to CA2687512A priority patent/CA2687512C/fr
Priority to JP2010510260A priority patent/JP2010527786A/ja
Priority to DK08767049.3T priority patent/DK2158166T3/da
Priority to US12/601,593 priority patent/US8377304B2/en
Publication of WO2008147296A1 publication Critical patent/WO2008147296A1/fr

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F11/00—Treatment of sludge; Devices therefor
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F1/00—Treatment of water, waste water, or sewage
    • C02F1/68—Treatment of water, waste water, or sewage by addition of specified substances, e.g. trace elements, for ameliorating potable water
    • C02F1/683—Treatment of water, waste water, or sewage by addition of specified substances, e.g. trace elements, for ameliorating potable water by addition of complex-forming compounds
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F11/00—Treatment of sludge; Devices therefor
    • C02F11/02—Biological treatment
    • C02F11/04—Anaerobic treatment; Production of methane by such processes
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00—Biological treatment of water, waste water, or sewage
    • C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
    • C02F3/342—Biological treatment of water, waste water, or sewage characterised by the microorganisms used characterised by the enzymes used
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P5/00—Preparation of hydrocarbons or halogenated hydrocarbons
    • C12P5/02—Preparation of hydrocarbons or halogenated hydrocarbons acyclic
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F1/00—Treatment of water, waste water, or sewage
    • C02F1/66—Treatment of water, waste water, or sewage by neutralisation; pH adjustment
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00—Biological treatment of water, waste water, or sewage
    • C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
    • Y02E50/00—Technologies for the production of fuel of non-fossil origin
    • Y02E50/30—Fuel from waste, e.g. synthetic alcohol or diesel

Definitions

  • the present invention relates to a novel method for treatment of sludge in water purification and the use thereof.
  • Background art The enzymatic hydrolysis of sludge has been investigated for the last two decades and a number of enzymes from different organisms have been reported to play an important role in a range of waste water treatment applications. Enzymes act on specific substances present in municipal sludge and therefore can change the characteristics of the waste. The sludge becomes more amenable to further treatment and bio-conversion to value-added products is facilitated. Enzymatic treatment of mixed sludge prior to the anaerobic digestion shows degradation of the sludge and lead to methane production. Enzymatic treatment of wastewater sludge has a potential contribution to the reduction of sludge volume without biomass generation.
  • Enzymes break down polymeric substances like proteins, polysaccharides and lipids through multi step processes. Initially, enzymes adsorb to the solid substrate and cleave small polymers that are loosely bound to the surface. The solubilisation of the more compact sludge core occurs at a lower rate dependent on the diffusion of the enzyme to surface active sites and core particles. Previous studies have shown that a combination of a protease, lipase and endo-glycanases accelerated solubilisation of municipal sludge. However, the enzymes became entrapped in the sludge. The entrapment decreased the enzymes action on the sludge but not on chromogenic soluble substrates present.
  • the present invention relates, in one aspect, to a method for treatment of sludge in water purification, characterized by the steps: a) providing an enzyme mixture comprising at least one enzyme capable of digesting natural polymeric materials; b) adding the enzyme mixture and at least one chelating reagent, in optional order or simultaneously, to an aqueous sludge suspension.
  • the present invention relates, in a further aspect, to the use of above mentioned method, in addition to conventional digestion used in water purification.
  • the present invention relates, in a yet further aspect, to the use of above-mentioned method, instead of conventional digestion used in water purification.
  • Fig. 1 depicts the COD (mg/l) released when using the chelating reagents citric acid (at 25 mM and 50 mM) and EDTA (at 25 mM and 50 mM) at different total enzyme doses (13.7 mg/g, 34.25 mg/g, and 54.8 mg/g) in comparison with in the absence of chelating reagents.
  • Fig. 2 depicts the ⁇ COD (mg/l) released when using enzymes together the chelating reagents citric acid (Cit A), EDTA, zeolite (Zeo), sodium fluoride
  • Fig. 3 depicts the activity (enzyme activity U/ml) of the enzymes proteases and celluiases in the solid and liquid phase in the presence of chelating reagents, Cit A and EDTA, at 25 mM and 50 mM as well as in the absence of any chelating reagent. The amount of enzymes in all samples was
  • the object of the present invention is to provide novel methods for treatment of sludge in water purification. It has been realized that adding a chelating agent in accordance with the method of the invention to purification methods enhances the digestion of the sludge. Furthermore, the chelating agent prevents enzymes from being trapped on the sludge surface and enzymatic solubilisation of sludge is improved.
  • the present invention would indeed be beneficial to the water purification industry for providing more efficient and cost effective processes. Therefore, enhancing the efficiency of the enzymes in water purification is the main focus of the present invention.
  • the at least one chelating reagent is chosen from the group consisting of citric acid (Cit A), ethylenediaminetetraacetic acid (EDTA), tartaric acid, and their salts, Zeolite A, sodium fluoride, sodium thiosulphate in combination with Zeolite A, sodium silicate, sodium silicate in combination with Zeolite A, and any combination of the above.
  • the chelating reagent may be a combination of two or more of the stated agents or any other chelating reagent not specifically mentioned herein. It is realized by a person skilled in the art that any other chelating reagent, not specifically mentioned herein, might be used in accordance with the invention as long as the desired result is achieved. However, chelating reagents containing phosphorous are not suitable because they increase the phosphorous load in the effluent.
  • the break-up of larger aggregates results in an increase of the specific surface area available for enzymatic hydrolysis.
  • the removal of poly cations from sludge decreases the degree of network but also changes the charges and polarity of the sludge surface.
  • electrostatic binding sites are a crucial indicator for bio-sorption of metals in wastewater treatment and in bioremediation. Sludge charge changes will effect all ionic interactions including the non-cation type. It has been shown that the lower the hydrolysis temperature the less negative groups are released.
  • the at least one chelating reagent is added to the sludge suspension in a total final concentration of about 0.1-200 mM (of the total sludge suspension), preferably added in a total final concentration of about 0.1 -75 mM, e.g. 1-75 mM (of the total sludge suspension).
  • the chelating agent can be added in a concentration of about 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, and 75 mM.
  • the chelating agent can be added in a concentration of about 0.1-50 mM, preferably 0.1-25 mM and more preferably 0.1-10 mM, e.g. 0.1-5 mM.
  • the chelating agent can be added in a concentration of about 1-50 mM, e.g. 1-25 mM.
  • an enzyme mixture capable of digesting natural polymeric materials. It is realized by a person skilled in the art that any type of enzyme, not specifically mentioned herein, may be used in the enzyme mixture as long as the desired effect is achieved.
  • the at least one enzyme in the enzyme mixture is chosen from glycosidic enzymes, lipases, proteases, oxidases, phytases.
  • the enzyme mixture may comprise one or more enzymes.
  • Other examples of enzymes that can be used are amylases, cellulases , xylanases, dextranases and glycanses.
  • the enzymes can be selected depending on the type and source of the sludge.
  • Preferred enzymes are proteases such as Alcalase (product name of commercially available protease) and lipases such as Lipolase (product name of commercially available lipase).
  • Alcalase is a protease that has a very broad specificity, namely high specificity for aromatic aminoacids such as Phe, Trp, and Tyr, acidic aminoacids GIu, sulfur-containing aminoacids such as Met, and aliphatic amino acids such as Leu and Ala .
  • aromatic aminoacids such as Phe, Trp, and Tyr
  • acidic aminoacids GIu acidic aminoacids
  • sulfur-containing aminoacids such as Met
  • aliphatic amino acids such as Leu and Ala .
  • protease shares specificities which are separately associated with a few proteases. The same is valid for the commercially av ailable Lipolase that has a broad substrate specificity. In other words, it promotes the hydrolysis of a wide variety of triglycerides.
  • the enzyme mixture is a combination of for instance at least two enzymes, e.g. the combination comprises for instance ⁇ -amylase and cellulase, laccase and lipolase, laccase and ⁇ -amylase, or any other combination of the above.
  • a further combination of enzymes is for instance a protease such as Alcalase and a lipase such as Lipolase.
  • the enzyme(s) in the enzyme mixture should be capable of digesting natural polymeric materials, i.e. have the ability to digest the present natural polymeric materials. This is an important feature of the enzyme(s), since otherwise the sludge would not be digested and/or broken down. In other words, the enzyme(s) is/are added to the water purification process for digesting natural polymeric materials. Thus, the enzymes are added in an effective amount to digest the present natural polymeric materials.
  • An advantage of the present invention is that the enzymatic dissolving process according to the present invention is much faster compared to a conventional enzymatic dissolving process.
  • the enzymatic dissolving process takes place for 1-12 hours, preferably 3-8 hours, more preferably 4-6 hours, for example 4 hours.
  • a conventional enzymatic dissolving process would require at least 12 hours.
  • One of the major sources of the organic matter in sludge are extracellular polymeric substances (EPS), found in two forms: soluble and bound EPS, depending on sludge source.
  • the soluble EPS can be extracted by centrifugation alone while bound EPS in floes of biomass requires additional treatments.
  • the major components in the EPS are polysaccharides and glycoconjugates.
  • the architecture of sludge floes is dependent on the interaction between the microbial polymers, acidic polysaccharides, glycoconjugates and lectin-like protein binding polysaccharides as well humates that are linked by poly-cations present in the sludge. It is sometimes required to adjust the pH of the sludge suspension in the method according to the invention to about 6-9, preferably 7-8, after addition of the at least one chelating reagent by adding an acid or base, for instance by adding HCI or NaOH or any other suitable base or acid.
  • the treatment takes place in any anaerobic digester, i.e. without the presence of oxygen.
  • the end product of an anaerobic process is methane, which can be recovered from the anaerobic process to obtain an additional product to be used in other applications.
  • methane which can be recovered from the anaerobic process to obtain an additional product to be used in other applications.
  • a value-added product is also obtained from the process.
  • a very important embodiment of the invention is the use of the method of the invention in an anaerobic environment, whereby a value added product such as methane can be separated and reused.
  • the mass ratio of the enzyme mixture:sludge dry solids is from about 0.1 - 140 mg/g dry solids, preferably 0.1-70 mg/g dry solids, e.g. 1 - 70 mg/g dry solids, more preferably 1-55, more preferably 1- 35 and more preferably 1-14 mg/g dry solids.
  • the mass ratio of enzyme mixture:sludge dry solids could for example be as low as preferably 0.1-5, 0.1-10 or 2-6 mg/g dry solids for economical reasons.
  • the mass ratio of the enzyme mixture:sludge suspension has been defined to the range of 0.1-140 mg/g dry solids.
  • the enzyme mixture could, in addition to enzymes, comprise other amounts of constituents such as water, or suitable organic or inorganic solvents or other components.
  • the enzyme mixture used in the method of the invention could for instance be a commercially available enzyme products containing the relevant enzyme(s) and amounts of solvent(s) and other components for these enzyme(s). It is naturally important that these other components and solvents do not disturb the important activity of the enzymes.
  • a non-limiting example of the invention is the case where a mass ratio of enzyme mixture: sludge dry solids is 13.7 mg/g, which means, in this non- limiting example, 13.7 mg of one or mixture of several commercial enzyme products per 1 g sludge dry solids.
  • the enzyme mixture used in the method is not to be limited to the above mentioned specific examples of commercially available enzyme products.
  • a person skilled in the art realizes the required amount of enzymes needed in the enzyme mixture to get an efficient degradation of sludge in view of the conditions of the process, i.e. temperature, type of sludge, and required efficiency and so on.
  • the method further comprises the step of adding at least one species of fermenting bacteria to the suspension, thereby fermenting the resulting sludge suspension obtained in step b) of the method of the invention.
  • it is also possible to further increase the degradation of the sludge by adding different fermenting microbes, e.g. fermenting bacteria, to the suspension.
  • the fermenting bacteria are for instance chosen from acidogenic bacteria, acetogenic bacteria, and methane producing bacteria.
  • the fermenting bacteria are chosen from the group consisting of Gluconobacter oxydans, Acetobacter sp., Acetogenium kivui, B. macerans, B. polymyxa, B. coagulans, Lactobacillus buchneri, Clostridium thermoaceticus, Clostridium lentocellum, Clostridium formicoaceticu, Clostridium thermocelium and Pseudomonas sp..
  • the methane producing bacteria are chosen from the group consisting of Methanosarcina barkeri, Methanosarcina mazeii, Methanosarcina soehngenii, Methanosarcina acetivorans, and Methanosaeta sp, and mixtures thereof.
  • the methane produced is separated from the sludge suspension.
  • an additional effect is achieved in accordance with this particular embodiment in the form of a value-added product.
  • natural polymeric materials that are digested by the enzymes are for instance proteins, polysaccharides, polyphenols, lignins, humates, fats, waxes, and mineral oils.
  • the phrase includes any other material or components, not specifically mentioned here, that are present in the sludge and are also effected by the present enzymes.
  • chelating reagent and "chelator” and “chelating agent” are used interchangeably. However, they all are to be interpreted in the same way as the generally known definition of chelating agent.
  • the temperature of the sludge suspension in the method of the invention is from about 4 0 C to about 9O 0 C, preferably from about 1O 0 C to about 80°C, and more preferably from about 3O 0 C to about 60 0 C, and even more preferably from 37°C to about 40 0 C.
  • the temperature is chosen on the basis of the present enzymes and their action on the natural polymeric materials. A skilled man in the art can determine the optimal conditions in order to achieve an efficient degradation.
  • the sludge suspension could be subjected to agitation in the range from above 0 to 200 rpm.
  • the agitation is beneficial from an efficiency point of view.
  • the enzymes are able to act more efficiently since the agitation causes the sludge to become more available.
  • the sludge is pre- concentrated, prior to the addition of enzymes, chelating agent and optionally bacteria, by gravitation or enhanced sedimentation to the range 10-80 g sludge solids per 1 I sludge suspension.
  • the total solid (TS) content was in the range of 2% and 4%.
  • Protease Alcalase 2.4L
  • lipase Lipolase 100L
  • glycanases dextranase (Dextranase PlusL), endo-xylanase (Pulpzyme HC), cellulase (Celluclast 1.5L), ⁇ -amylase (Termamyl 300L) were a gift from Novozyme A/S, Denmark.
  • the enzyme activity was measured in both liquid and solid part of sludge using soluble chromogenic substrates accordingly to the assay procedures from Megazyme, Ireland (Megazyme).
  • the following substrates were used (i) for protease, AZO-Casein (2% solution in 0.1 M phosphate buffer, pH 7.0), (ii) for cellulase, AZO-CM-Cellulose (2% solution in 0.1 M sodium acetate buffer, pH 4.5).
  • One enzyme unit of activity is defined as the amount of the enzyme required to hydrolyze one micromole of soluble substrate per minute under standard assay conditions (i: pH 7.0 or ii: 4.5 and 45°C).
  • Enzymatic treatments were performed in 1L reactors under controlled temperature conditions (45 + 1 0 C) and the pH was constant during the process (pH 7).
  • Chelating agents for example citric acid (Cit A) or EDTA were added to sludge as powder to a final concentration of 25 mM or 50 mM. Addition of 50 mM EDTA or Cit A caused the pH to decrease to 5.5 and 3.3, respectively. Before enzymes were added the pH of sludge was adjusted to 7 using 1M NaOH.
  • 60 mg of the enzyme mixture A contained: 12 mg of each of the following enzyme preparations from Novozyme A/S: Termamyl 300 L, Type DX (amylase) Declared activity: 300 KNU/g (37 0 C pH.5.6), Lipolase 100 L (lipase) 100 KLU/g (3O 0 C, pH 7.0), Celluclast 1.5 L (cellulase) 700 EGU/g (3O 0 C pH.5.6), Pulpzyme HC (endo-Xylanase) 1000 AXU/g; and Dextranase Plus L(dextranase), and the enzymes were suspended in 1.2 mg of PPG 4 and 0.12 mg of FAE.
  • 8.5 mg of enzyme mixture B contained 8.5 mg of enzyme preparation Alcalase 2.4 LFG (protease) 2.4 AU-A/g (37 0 C pH.8.5).

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Abstract

La présente invention concerne un procédé de traitement d'une boue dans la purification de l'eau, le procédé comprenant les étapes consistant à : fournir un mélange d'enzymes comprenant au moins une enzyme capable de digérer les substances polymères naturelles; et ajouter le mélange enzymatique et au moins un réactif chélatant, dans un ordre éventuel ou simultanément, à une suspension de boue aqueuse. L'agent chélatant est choisi dans le groupe comprenant l'acide citrique et ses sels.
PCT/SE2008/000376 2007-05-31 2008-05-30 Procédé de traitement enzymatique d'une boue dans la purification de l'eau WO2008147296A1 (fr)

Priority Applications (8)

Application Number Priority Date Filing Date Title
AU2008257660A AU2008257660A1 (en) 2007-05-31 2008-05-30 A method for enzymatic treatment of sludge in water purification
CN2008800178774A CN101679089B (zh) 2007-05-31 2008-05-30 一种用于在水净化中酶处理污泥的方法
EP08767049A EP2158166B1 (fr) 2007-05-31 2008-05-30 Procédé de traitement enzymatique d'une boue dans la purification de l'eau
BRPI0810931-1A2A BRPI0810931A2 (pt) 2007-05-31 2008-05-30 Método para o tratamento da lama na purificação da água
CA2687512A CA2687512C (fr) 2007-05-31 2008-05-30 Procede de traitement enzymatique d'une boue dans la purification de l'eau
JP2010510260A JP2010527786A (ja) 2007-05-31 2008-05-30 浄水法における酵素的なスラッジ処理方法
DK08767049.3T DK2158166T3 (da) 2007-05-31 2008-05-30 Fremgangsmåde til enzymbehandling af slam ved vandrensning
US12/601,593 US8377304B2 (en) 2007-05-31 2008-05-30 Method for treatment of sludge in water purification

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
SE0701378-2 2007-05-31
SE0701378A SE531167C2 (sv) 2007-05-31 2007-05-31 Förfarande för behandling av slam vid vattenrening

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WO2008147296A1 true WO2008147296A1 (fr) 2008-12-04

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US (1) US8377304B2 (fr)
EP (1) EP2158166B1 (fr)
JP (1) JP2010527786A (fr)
KR (1) KR20100017936A (fr)
CN (1) CN101679089B (fr)
AU (1) AU2008257660A1 (fr)
BR (1) BRPI0810931A2 (fr)
CA (1) CA2687512C (fr)
DK (1) DK2158166T3 (fr)
RU (1) RU2009146272A (fr)
SE (1) SE531167C2 (fr)
WO (1) WO2008147296A1 (fr)

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US9388369B2 (en) 2010-08-20 2016-07-12 Ecolab Usa Inc. Wash water maintenance for sustainable practices
CN108046557A (zh) * 2017-09-20 2018-05-18 同济大学 促进含磷酸盐沉淀的污泥在室温条件下释磷与产气的方法
US10047339B2 (en) 2011-04-04 2018-08-14 Drylet, Llc Composition and method for delivery of living cells in a dry mode having a surface layer
WO2018160567A1 (fr) * 2017-02-28 2018-09-07 Drylet, Llc Systèmes, procédés et appareil pour augmenter la qualité d'effluent et de biosolides d'eaux usées
US10316312B2 (en) 2009-10-05 2019-06-11 Drylet, Llc Composition and method for delivery of microorganisms in a dry mode in porous structure

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US10563113B2 (en) * 2016-01-05 2020-02-18 Saudi Arabian Oil Company Removal of barite weighted mud
CN106007313A (zh) * 2016-07-28 2016-10-12 北京赛富威环境工程技术有限公司 一种污泥的分级减量处理方法
FI20165978L (fi) * 2016-12-16 2018-06-17 Kemira Oyj Menetelmä veden poistamiseksi lietteestä
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CN110709370A (zh) * 2017-02-28 2020-01-17 德莱特有限公司 用于提高废水流出物和生物固体的质量的系统、方法和设备
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CN108046557A (zh) * 2017-09-20 2018-05-18 同济大学 促进含磷酸盐沉淀的污泥在室温条件下释磷与产气的方法
CN108046557B (zh) * 2017-09-20 2021-02-09 同济大学 促进含磷酸盐沉淀的污泥在室温条件下释磷与产气的方法

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CN101679089A (zh) 2010-03-24
DK2158166T3 (da) 2013-03-18
BRPI0810931A2 (pt) 2014-12-23
US8377304B2 (en) 2013-02-19
RU2009146272A (ru) 2011-07-10
AU2008257660A1 (en) 2008-12-04
CA2687512C (fr) 2015-09-01
KR20100017936A (ko) 2010-02-16
EP2158166A1 (fr) 2010-03-03
EP2158166A4 (fr) 2010-08-25
EP2158166B1 (fr) 2012-12-05
CA2687512A1 (fr) 2008-12-04
US20100200499A1 (en) 2010-08-12
JP2010527786A (ja) 2010-08-19
SE0701378L (sv) 2008-12-01
CN101679089B (zh) 2012-08-08
SE531167C2 (sv) 2009-01-07

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